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Super-resolution Fluorescence Microscopy

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.

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Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.

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Confocal Fluorescence Microscopy

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Updated: Jul 3, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

On the fluorescence from integrating spheres.

Ping-Shine Shaw¹, Zhigang Li

¹National Institute of Standards and Technology, Gaithersburg, Maryland 20899, USA. shaw@nist.gov

|July 22, 2008

Summary

UV-induced fluorescence in integrating spheres is amplified by hydrocarbon contamination and multiple reflections. This study presents a theory explaining fluorescence dependence on material reflectance and sphere geometry, revealing significant signal gains.

Area of Science:

Optical Engineering
Materials Science

Background:

Integrating spheres are crucial optical components.
UV light can induce fluorescence in diffusing materials.
Hydrocarbon contamination is a known issue affecting optical performance.

Purpose of the Study:

To develop a theoretical model for UV-induced fluorescence in integrating spheres.
To investigate the impact of material properties and geometry on fluorescence.
To understand the amplification of fluorescence due to multiple reflections.

Main Methods:

Theoretical modeling from first principles.
Analysis of fluorescence dependence on reflectance and geometry.
Comparison with single-irradiation fluorescence.

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Last Updated: Jul 3, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope

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Published on: October 28, 2018

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

Single Molecule Fluorescence Microscopy on Planar Supported Bilayers

Published on: October 31, 2015

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

Published on: February 8, 2015

Main Results:

Fluorescence is strongly dependent on material reflectance at excitation and emission wavelengths.
Integrating sphere geometry significantly influences fluorescence intensity.
Multiple reflections can amplify fluorescence by over an order of magnitude.

Conclusions:

A theoretical framework for understanding integrating sphere fluorescence has been established.
Hydrocarbon contamination can lead to significant, amplified UV-induced fluorescence.
This phenomenon has implications for the calibration and application of integrating spheres.